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COMPANY CONFIDENTIAL - FOR INTERNAL USE ONLY
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ACQUITY UPLC® H-Class System and H-Class Bio System with TUV, PDA, ELS, FLR, or RI Detectors
Installation Manual
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Notice The information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This checklist is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with or arising from the use of this document. ©2013 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA. ALL RIGHTS RESERVED. THIS BOOK OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER. Waters, ACQUITY UPLC, and Connections INSIGHT are registered trademarks of Waters Corporation. THE SCIENCE OF WHAT’S POSSIBLE, Empower, and MassLynx are trademarks of Waters Corporation. All other trademarks are the sole property of their respective owners.
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Table of Contents
Preparing mobile phases and wash solvent for the performance tests . . . . . . . 4 Tools/materials required . . . . . . . . . . . . . . . . . . . . Solvent and glassware considerations . . . . . . . . . . . Preparing the mobile phases . . . . . . . . . . . . . . . . . Preparing the wash and purge solvent (for TUV, PDA,
......................... ......................... ......................... and ELS detectors) . . . . . . . . . . .
Installing the ACQUITY UPLC H-Class System modules
.. .. .. ..
.4 4 .5 5
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Before you begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tools/materials/software required. . . . . . . . . . . . . . . . . . . . . . . . . . . Example system configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . Installing the Quaternary Solvent Manager (QSM) . . . . . . . . . . . . . . . Installing the Sample Manager - Flow Through Needle (SM - FTN) . . . . Installing an optional Column Heater (CH-A) . . . . . . . . . . . . . . . . . . . Installing an optional Column Manager-A / Auxiliary Column Manager-A Installing the optical detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Installing the solvent tray module. . . . . . . . . . . . . . . . . . . . . . . . . . .
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Making the fluidic connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Making the power and electronic connections . . . . . . . . . . . . . . . . . . . . . . . . . 21 Powering-on the system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 Restoring the startup project and configuring the system (Empower-based systems) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 Configuring the system (MassLynx-based systems) . . . . . . . . . . . . . . . . . . . . 24 Running the performance tests for systems with a TUV, PDA, ELS, FLR, or RI detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 Test solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tools/materials required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preparing samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . For TUV and PDA detectors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . For ELS detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . For FLR detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . For RI Detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preparing diluent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preparing the system precision test sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preparing the gradient performance test sample . . . . . . . . . . . . . . . . . . . . . . . . Preparing for the system performance test . . . . . . . . . . . . . . . . . . . . . . . . . . . . Running maintenance tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Running a system leak test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Connecting and equilibrating the column . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Running the system precision test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Checking the method parameters (TUV, PDA, ELS, FLR) . . . . . . . . . . . . . . . . Creating the method parameters (RI only) . . . . . . . . . . . . . . . . . . . . . . . . . . System precision test results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Running the gradient performance test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Gradient performance test results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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Preparing mobile phases and wash solvent for the performance tests CAUTION:
TO PREVENT CONTAMINATION, WEAR CLEAN, CHEMICAL-RESISTANT, POWDER-FREE
GLOVES.
Tools/materials required • • • • •
100 mL volumetric flask 100 mL graduated cylinder 1 L reservoir bottle Pipette Fisher Optima HPLC-grade acetonitrile (TUV, PDA, ELS, FLR)
•
Waters RI Detector Qualification Mobile Phase (part number 700002419 for RI only)
• • • •
Baker HPLC-grade water Solvent Clarification Kit (part number WAT200543) General-purpose laboratory vacuum pump ACQUITY UPLC BEH C18 (2.1 x 50 mm, 1.7 µm, part number 186002350)
NOTE:
Waters recommends purchasing the pre-mixed RI Detector Qualification Mobile Phase. However, it can be prepared by pre-mixing HPLC-grade 20.9% Methanol/79.1% ASTM Type1 Reagent Water, weight/weight.
Solvent and glassware considerations Due to the increased sensitivity that both the ACQUITY UPLC H-Class System and new series of detectors offer, all solvents including water and additives must be of the highest chemical purity (HPLC-grade or better). Failure to use HPLC-grade solvents will result in high background contamination, low signal-to-noise, and loss of sensitivity. Waters recommends using Fisher Optima HPLC-grade acetonitrile (TUV, PDA, ELS, FLR) or Waters RI Detector Qualification Mobile Phase (RI only) and Baker HPLC-grade water. Glassware (e.g. solvent bottles) must not be washed with detergents or washed with other general glassware to prevent contamination. The glassware must be rinsed with the high purity solvents to be used. NOTE:
For further detail on controlling contamination, refer to the document Controlling Contamination in Ultra Performance LC™/MS and HPLC/MS Systems, part number 715001307, located in the Waters Support Center on the Web (www.waters.com).
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Waters recommends that mobile phases and wash solvents be mixed as shown in Table 1 (TUV, PDA, ELS, FLR). Table 1: Mobile Phase and Solvent Concentrations (TUV, PDA, ELS, FLR) Mobile Phase and Wash Solvents
Concentration
Mobile Phase A
100% water
Mobile Phase B
100% acetonitrile
Mobile Phase C
100% water
Mobile Phase D
100% acetonitrile
Purge solvent
90:10 water/acetonitrile
Seal Wash
90:10 water/acetonitrile
Wash solvent
90:10 water/acetonitrile (for TUV, PDA, ELS) 100% acetonitrile (for FLR)
Preparing the mobile phases 1. Prepare a mobile phase of 100% HPLC-grade water (TUV, PDA, ELS, FLR only): a. Pour approximately 1 L of HPLC-grade water into a 1-L reservoir bottle. b. Label the reservoir bottle: “100% water mobile phase”. 2. Prepare a mobile phase of 100% acetonitrile (TUV, PDA, ELS, FLR only): a. Pour approximately 1 L of filtered acetonitrile into a 1-L reservoir bottle. b. Label the reservoir bottle: “100% acetonitrile mobile phase”. 3. Prepare a mobile phase of 100% Waters RI Detector Qualification Mobile Phase (RI only): a. Pour approximately 1 L of filtered Waters RI Detector Qualification Mobile Phase into a 1-L reservoir bottle. b. Label the reservoir bottle: “100% Waters RI Detector Qualification Mobile Phase”. NOTE: For the RI detector, all solvent lines (A, B, C, D) and the purge, seal wash, and wash solvent lines will all be put into the reservoir labelled “100% Waters RI Detector Qualification Mobile Phase”.
Preparing the wash and purge solvent (for TUV, PDA, and ELS detectors) NOTE:
The wash solvent for FLR detectors uses the “100% acetonitrile mobile phase”. The wash and purge solvents for RI detectors use the “100% Waters RI Detector Qualification Mobile Phase”.
1. Pour 90 mL of filtered HPLC-grade water into a reservoir bottle. 2. Pour 10 mL of filtered acetonitrile into the same reservoir bottle. 3. Cap the reservoir bottle and mix well. 4. Label the reservoir bottle “90% Water/10% Acetonitrile”.
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Installing the ACQUITY UPLC H-Class System modules Before you begin • If the system includes an optional Flex Cart, assemble the cart according to the instructions in Assembling the Flex Cart (P/N 715001019). • If the system includes multiple detectors, refer to Configuring the ACQUITY UPLC System Components (P/N 715001701) for configuration details. • If the system includes a Column Manager-A (CM-A) or Auxiliary Column Manager (CM-Aux), refer to Installing an H-Class System Column Manager-A / Auxiliary Column Manager (P/N 715002684) for detailed install steps. CAUTION:
TO AVOID INJURY, AT LEAST TWO PEOPLE MUST LIFT THE SAMPLE ORGANIZER, QUATERNARY SOLVENT MANAGER, AND THE SAMPLE MANAGER-FTN.
CAUTION:
TO AVOID FINGER INJURY, USE CAUTION WHEN PLACING THE QUATERNARY SOLVENT MANAGER AND SAMPLE MANAGER-FTN MODULES ON THE BENCH TOP OR FLEX CART.
Tools/materials/software required • • • • • • • •
ACQUITY UPLC H-Class System Start-up Kit Powder-free gloves PEEK tubing cutter 1/4-inch open-end wrench 5/8-inch open-end wrench Phillips head screwdriver EMP_HCLASS_startup project (for Empower-based systems) ML_HCLASS_startup project (for MassLynx-based systems)
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Example system configurations There are several possible configurations for your ACQUITY UPLC H-Class System. Many configurations use a single-stack footprint; dual detector with a CM-A/CM-Aux configuration requires a dual-stack footprint; some configurations can require a triple-stack footprint. Figure 1 shows example configurations for various single-, dual-, and triple-stack footprints.
Extension bracket
PDA TUV
TUV
CH-A
CH-A
SM-FTN QSM
Single Stack
CM-Aux CM-A
CH-A
SM-FTN
SM-FTN
SM-FTN
QSM
QSM
QSM
CH-30A
Single Stack (with CH-30A)
Single Stack (CM-A and CM-Aux)
CM-A SM-FTN
TUV
SM-FTN
Dual Stack
Dual Stack
(dual detectors with CH-A and additional SM-FTN spacing)
(dual detectors with additional SM-FTN and CM-A spacing)
QSM
CH-A
CH-A
CM-Aux
SQD
CM-A
SM-FTN
SQD
SM-FTN QSM
QSM
CM-Aux
Dual Stack
Dual Stack (CM-A and two CM-Aux)
Dual Stack (with CH-30A)
(with additional ELS drain tube spacing)
ELS PDA SM-FTN QSM
CM-Aux CM-A CM-Aux
Dual Stack (With additional SM-FTN spacing)
PDA SM-FTN QSM
SQD
QSM
ELS
ELS PDA
TUV
CM-Aux
SQD
CM-A CM-Aux
Triple Stack (With additional SM-FTN and CM-A spacing)
Figure 1 - Configuration examples (Front View)
CH-30A
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Installing the Quaternary Solvent Manager (QSM) 1. Carefully place the QSM onto the bench top or optional flex cart.
Installing the Sample Manager - Flow Through Needle (SM - FTN) 1. Carefully place the SM-FTN on top of the QSM. 2. Follow step 3 through step 5 only if your system includes a CH-A. 3. On the back of the SM-FTN, remove the center screw that secures the top cover to the unit. You will replace the screw with another from the CH-A startup kit after installing the CH-A.
Figure 2 - Center screw removed 4. From the CH-A startup kit, locate the CH-A-to-SM-FTN bracket (P/N 400004207) and screw (P/N 410000419). See Figure 3. 5. Align the hole in the bracket with the threaded hole on the front-left of the SM-FTN top cover, then secure it in place using a T20 Torx driver (Figure 3).
Cutout for CH-A “foot”
Figure 3 - Securing the CH-A-to-SM-FTN bracket to the SM-FTN
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Installing an optional Column Heater (CH-A) NOTE:
Skip this section if your system includes an optional Column Manager-A (CM-A) or CM-A with Auxiliary Column Manager (CM-Aux).
1. Carefully place the CH-A on top of the SM-FTN. 2. Align the CH-A with the SM-FTN as follows: • the two studs on the back of the SM-FTN align with and extend through the two slots in the back plate of the CH-A (Figure 4) • the front-left “foot” of the CH-A is seated in the cutout of the CH-A-to-SM-FTN bracket (Figure 3) • the hole in the center of the CH-A back plate is aligned with the threaded screw hole in the SM-FTN
CH-A
Slot Stud
SM-FTN
Figure 4 - Position on the SM-FTN 3. Using the blue Phillips head screw with nylon washer from the CH-A startup kit, secure the center of the CH-A back plate to the SM-FTN.
Installing an optional Column Manager-A / Auxiliary Column Manager-A NOTE:
Skip this section if your system includes a Column Heater-A (CH-A).
1. Position the CM-A (and if applicable one or two CM-Aux modules) as defined in the Installing an H-Class System Column Manager-A / Auxiliary Column Manager (P/N 715002684). 2. After positioning all system components, refer to this document to: a. Make the fluidic connections for the QSM, SM-FTN, and any detectors. b. Make the power and electrical connections for all components. c. Power on all system components and confirm communications. d. Set the desired CM-A / CM-Aux column configuration, and confirm that the CM-A valve positions are set to “waste,” as described in Installing an H-Class System Column Manager-A / Auxiliary Column Manager (P/N 715002684). e. Plumb the CM-A / CM-Aux module(s) in preparation for the system performance test, as described in Installing an H-Class System Column Manager-A / Auxiliary Column Manager (P/N 715002684).
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Installing the optical detector 1. Carefully place the TUV, PDA, ELS, FLR, or RI into position on the stack. NOTE: If the system includes a CM-A with one or two CM-Aux modules, refer to the ACQUITY H-Class System Column Manager/Auxiliary Column Manager Installation Instructions” (P/N 715002684) for positioning details. For the TUV, PDA, ELS, and FLR detectors, if the system includes multiple detectors, refer to “Configuring the ACQUITY UPLC System Components” (P/N 715001701) for positioning details. For the RI detector, if the system includes multiple detectors, refer to the documents “RI Detector Aux Drip Tray Kit Installation” (P/N 715004088) and “Installing the ACQUITY RI Detector” (P/N 715004089).
Installing the solvent tray module 1. Place the solvent tray module on the stack, as appropriate. NOTE: For solvent-bottle accessibility, provide an extra 10-inch (25.4-cm) clearance above the top of the solvent tray. NOTE: If the system includes a CM-A with one or two CM-Aux modules, refer to the ACQUITY H-Class System Column Manager/Auxiliary Column Manager Installation Instructions” (P/N 715002684) for positioning details. If the system includes multiple detectors, refer to “Configuring the ACQUITY UPLC System Components” (P/N 715001701) for positioning details.
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Making the fluidic connections CAUTION:
TO PREVENT CONTAMINATION, WEAR CLEAN, CHEMICAL-RESISTANT, POWDER-FREE
GLOVES.
WARNING: TO AVOID RELEASING SOLVENT VAPORS INTO THE ROOM, ROUTE THE DEGASSER VENT TUBE INTO A FUME HOOD OR OTHER SUITABLE EXHAUST SYSTEM, OR TO A SUITABLE WASTE CONTAINER, ENSURING THE END OF THE TUBING IS ABOVE THE FLUID LEVEL. CAUTION:
TO AVOID FLUID BACKUP, ENSURE THAT THE WASTE CONTAINER IS BELOW THE STACK, ENSURE THERE ARE NO CRIMPS OR BENDS IN THE WASTE LINES, AND ENSURE WASTE AND DEGASSER LINE ENDS ARE NOT IMMERSED IN WASTE FLUID.
1. Release the degasser vent line from its holding clip on the QSM.
Clip
Figure 5 - Unclipping the degasser vent line 2. Locate the solvent tubing labeled “Sample Manager Wash” (or “Sample Manager 2”) and remove its solvent filter.
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3. Route the “Sample Manager Wash” solvent tubing through the tubing management channel in the SM-FTN, then reattach the solvent filter and insert the tubing into the reservoir labeled “90% water/10% acetonitrile” (for TUV, PDA, and ELS detectors), or into the reservoir labeled “100% acetonitrile” (for FLR detectors), or into the reservoir labelled “100% Waters RI Detector Qualification Mobile Phase” (for RI detectors).
Channel
Sample Manager Wash tubing
Figure 6 - Routing the Sample Manager Wash tubing 4. Route the A, B, C, and D solvent tubing through the outside of the channel.
Figure 7 - Routing the solvent tubing 5. Insert these solvent lines into their appropriate reservoirs: a. Insert Solvent A and Solvent C tubing into the 100% water reservoir (TUV, PDA, ELS, FLR). b. Insert the Seal Wash In tubing into the 100% water reservoir (TUV, PDA, ELS, FLR). c. Insert Solvent B and Solvent D tubing into the 100% acetonitrile reservoir (TUV, PDA, ELS, FLR).
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NOTE: For the RI detector, insert the solvent lines (A, B, C, D) into the reservoir labelled “100% Waters RI Detector Qualification Mobile Phase”. 6. Connect the “Sample Manager Purge” (or “Sample Manager 1”) tubing to either of the two SM SYRINGE ports on the QSM.
Figure 8 - Sample Manger Purge tubing connection to the QSM 7. Route the other end of the “Sample Manager Purge” solvent tubing through the channel and insert it into the 90% water/10% acetonitrile reservoir (TUV, PDA, ELS, FLR). NOTE: For the RI detector, insert the purge solvent line into the reservoir labelled “100% Waters RI Detector Qualification Mobile Phase”. 8. Unfasten the taped syringe waste and purge solvent tubing from the inside of the SM-FTN door.
Syringe waste tubing
Purge solvent tubing
Figure 9 - Syringe waste and purge solvent tubing
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9. Connect the Purge Solvent tubing to the available SM SYRINGE port on the QSM.
Purge solvent tubing
Figure 10 - Purge Solvent tubing connection to QSM 10. Route the syringe waste line behind the stainless steel clip on the QSM.
Syringe waste line
Clip
Figure 11 - Syringe waste line routing
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11. Use a tubing cutter to cut the syringe waste line to allow connection to its waste port on the QSM drip tray.
Syringe waste line
Figure 12 - Syringe waste port connection 12. Connect the convoluted waste tubing to the SM-FTN waste port, then route it through the hole in the SM-FTN drip tray.
SM-FTN waste tubing
Figure 13 - SM-FTN waste tube routing
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13. Route the SM-FTN waste tubing behind the stainless steel clip on the QSM, then connect it to the waste port on the QSM drip tray.
Waste port
Figure 14 - SM-FTN waste port connection 14. Route the degasser vent tube as shown in Figure 15, then on to a suitable container or exhaust system.
Degasser waste tube
Figure 15 - Degasser waste tube routing WARNING: TO AVOID RELEASING SOLVENT VAPORS INTO THE ROOM, ROUTE THE DEGASSER VENT TUBE INTO A FUME HOOD OR OTHER SUITABLE EXHAUST SYSTEM, OR TO A SUITABLE WASTE CONTAINER, ENSURING THE END OF THE TUBING IS ABOVE THE FLUID LEVEL. 15. Connect a section of Tygon tubing from the QSM waste outlet to a waste reservoir.
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16. Remove the long compression screw and ferrules from the stainless steel PUMP OUTLET tube, then route the end of the tube through the channel in the SM-FTN. Long compression screw
Short compression screw
Figure 16 - Pump Outlet tube routing 17. Reattach the long compression screw and ferrules to the PUMP OUTLET TUBE, then secure to port 5 on the inject valve (finger-tight plus 3/4-turn using the 1/4-inch openend wrench).
Port 5 (long compression screw)
Figure 17 - Pump Outlet tube inject valve connection
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18. Remove the inline filter from its clip on the QSM, then attach the other end of the PUMP OUTLET tube to the inline filter using the 5/8- and 1/4-inch open-end wrenches.
Figure 18 - Pump Outlet tube to inline filter connection 19. Install the SM-FTN drip tray curtain (P/N 415001552) onto the bottom of the lower drip tray as shown in Figure 19.
Figure 19 - Installing the SM-FTN drip tray curtain
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20. Connect the active pre-heater (APH) assembly to port 6 of the inject valve, then carefully apply a bend to the tube, making sure the tube exits through the tubing channel.
APH tube
Figure 20 - APH tube inject valve connection 21. Refer to Figure 21 to identify the style of Active Pre-Heater (APH) finger-tight fittings that ship with your system. NOTE:There are two styles of finger-tight fittings that ship: modules ship with a single-threaded style fitting that uses a collet; later modules ship with a dualthreaded fitting and a stainless steel cap nut. Finger-tight fitting (single thread)
Ferrule
Collet
Stainless cap nut
Dual-threaded finger-tight fitting
Ferrule
Figure 21 - Two styles of finger-tight APH fittings 22. Do one of the following: • If your shipment includes a single-threaded fitting with collet, follow step 23 to install the V-detail union (Start-up kit) to the outlet of the Active Pre-Heater (APH). • If your shipment includes a dual-threaded fitting with cap nut, follow step 24 to install the V-detail union (Start-up kit) to the outlet of the Active Pre-Heater (APH).
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23. For APH assemblies that use a single-threaded finger tight fitting (Figure 21), attach the V-detail union (Start-up kit) to the outlet of the APH assembly that is installed in the topright column compartment position: CAUTION: TO AVOID DAMAGING THE APH TUBING, DO NOT OVER-TIGHTEN THE FITTING. OVERTIGHTENING THE FITTING COULD SEPARATE THE TUBING COLLAR FROM THE INTERNAL WELD CAUSING LEAKS. a. Remove the APH assembly from the retainer in the column compartment. b. Before connecting the union to the fitting, make sure the ferrule is near the end of the tubing and the collet touches the back of the ferrule. c. Push the tubing into the union until it stops. d. Without placing any strain on the APH cartridge assembly, carefully screw the union into the finger-tight fitting. e. Finger-tighten the fitting until snug, then tighten an additional 1/4-turn. f. Go to step 25. 24. For APH assemblies that use a dual-threaded finger tight fitting (Figure 21), attach the V-detail union (Start-up kit) to the outlet of the APH assembly that is installed in the topright column compartment position: a. Remove the APH assembly from the retainer in the column compartment. b. Loosen the stainless-steel cap nut from the finger-tight fitting. c. Insert the stainless-steel tubing with ferrule and fitting into the union. Make sure the stainless-steel tubing protrudes beyond the ferrule. d. Seat the ferrule by securely finger-tightening the fitting into the union. Do not engage the stainless-steel cap nut when seating the ferrule. e. Complete the connection by finger-tightening the stainless-steel cap nut onto the fitting. 25. Connect a waste line to the union outlet. 26. Plug the APH assembly with attached union and waste tubing into the column compartment. 27. Connect the waste line to a waste container. NOTE: You will connect the column after priming and flushing the system in preparation for the system performance tests. 28. Connect Tygon tubing from the back of the solvent module tray to waste.
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Making the power and electronic connections 1. Connect Ethernet cables from the following instruments to the 6-port Ethernet block on the SM-FTN: • Computer • QSM • CM-A (if applicable) • Detector(s) 2. If applicable, connect the 25-pin D-shell connector from CH-A to the QSP1 port on the SM-FTN. 3. Connect all power cords. 4. Refer to Figure 22 and Figure 23, as necessary.
Figure 22 - Power and electronic connections (for system with a CH-A)
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CM-Aux
To upper CM-Aux
To lower CM-Aux
CM-A Not required
CM-Aux
Figure 23 - Power and electronic connections (for system with a CM-A and two CM-Aux modules)
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5. Install the two leak sensors on the SM-FTN and one on the QSM. Figure 24 shows the upper and lower SM-FTN sensor connection details.
Upper leak sensor
Lower leak sensor
Figure 24 - SM-FTN leak sensor connections
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Powering-on the system 1. Power on the PC, then install Empower or MassLynx. Ensure that the appropriate service packs/SCNs/software builds are installed. 2. Starting with the SM-FTN, power on all ACQUITY H-Class instruments (booting can take several minutes). 3. After a successful boot, launch Empower or MassLynx and ensure communication with the system instruments. 4. Do one of the following: • If the system is Empower-based, go to Restoring the startup project and configuring the system (Empower-based systems). • If the system is MassLynx-based, go to Configuring the system (MassLynx-based systems).
Restoring the startup project and configuring the system (Empowerbased systems) For Empower-based systems, restore the EMP_HCLASS_startup project to the system PC before configuring the system. 1. Download the Empower EMP_HCLASS_startup project from the Waters Service Knowledge Center: a. Click the Knwl Ctr - Svc & Support tab. b. Click the Support Information folder in the Detailed Navigation area. c. Click Reference Information to open the Service Engineer Information window. d. From the Systems pull-down menu, select ACQUITY UPLC H-Class. e. Click Reference Information. f. Click System-level Information, then click the ACQUITY H-Class System with TUV, PDA, ELS, FLR, or RI Detectors Installation Manual. g. Download the Empower startup project. 2. From the Projects view of the Configuration Manager, right-click and select Restore Project. 3. Navigate to the EMP_HCLASS_startup project, then use the Restore Projects Wizard to restore the project. 4. Configure the system from Empower Pro.
Configuring the system (MassLynx-based systems) 1. Download the MassLynx startup project file from the Waters Service Knowledge Center. (Follow steps 1a through 1g, above, to download the MassLynx project). 2. Copy the downloaded startup project file to the MassLynx folder. 3. From Console, select the ACQUITY H-CLASS System, then configure the system.
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Running the performance tests for systems with a TUV, PDA, ELS, FLR, or RI detector Test solutions Table 2: Test Solutions Part Number
a
Description
Quantity
700002669
Waters ACQUITY UPLC System Absorbance Startup Solution (TUV and PDA)
1
700002848
Waters ACQUITY UPLC System ELSD Test Solution (ELSD)
1
700003694
Waters ACQUITY UPLC System FLR Test Solution (FLR)
1
700001713
Waters RI Detector OQ/PQ Solution
1
700002419
Waters RI Detector Qualification Mobile Phasea
1
Waters recommends purchasing the pre-mixed RI Detector Qualification Mobile Phase. However, it can be prepared by pre-mixing HPLC-grade 20.9% Methanol/79.1% ASTM Type1 Reagent Water, weight/weight.
Tools/materials required • Union from the Start-up kit • Pin plug • Business card (for calibrating the z axis)
Preparing samples For TUV and PDA detectors Use Waters ACQUITY UPLC Systems Absorbance Start-Up Solution (TUV and PDA only). 700002669 - 2 ampules of standard solutions containing: • Ampule# 700002642-6:
0.100 mg/mL caffeine in HPLC-grade water/acetonitrile (90:10)
• Ampule# 700002642-11:
4.0 µg/mL 2-acetylfuran in HPLC-grade water/acetonitrile (90:10) 4.0 µg/mL acetanilide in HPLC-grade water/acetonitrile (90:10) 4.0 µg/mL acetophenone in HPLC-grade water/acetonitrile (90:10) 4.0 µg/mL propiophenone in HPLC-grade water/acetonitrile (90:10) 4.0 µg/mL butylparaben in HPLC-grade water/acetonitrile (90:10) 4.0 µg/mL benzophenone in HPLC-grade water/acetonitrile (90:10)
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4.0 µg/mL valerophenone in HPLC-grade water/acetonitrile (90:10)
For ELS detectors Use Waters ACQUITY UPLC Systems ELSD Test Solution (ELSD only). 700002848 - 1 ampule of 10 mL standard solution containing: • 0.16 mg/mL caffeine in HPLC-grade water/acetonitrile (90:10)
For FLR detectors Use Waters ACQUITY UPLC Systems FLR Test Solution (FLR only). 700003694 - 1 ampule of 10 mL standard solution containing:
• 5 pg/µL anthracene in HPLC-grade water/acetonitrile (80:20)
For RI Detectors Use Waters RI Detector OQ/PQ Solution (RI only). 700001713 - 1 ampule of 10 mL standard solution containing: • 2.0 g/L caffeine in HPLC-grade water/methanol (81%:19% weight:weight or 77:23 volume:volume)
Preparing diluent 1. Fill an appropriate sized sample vial with 10:90 Acetonitrile/Water (TUV, PDA, ELS, FLR) or RI Detector Qualification Mobile Phase (79.1%:20.9% water/methanol; RI only). 2. Label the vial “Diluent”.
Preparing the system precision test sample 1. Fill an appropriate sized sample vial with caffeine test solution from ampule # 700002642-6 (TUV and PDA only) or ampule # 700002848 (ELSD only), or ampule # 700001713 (RI only), or with the anthracene test solution from ampule # 700003694 (FLR only). 2. Label the vial as System Precision Test.
Preparing the gradient performance test sample 1. Fill an appropriate sized sample vial with the 7-component mix test solution from ampule # 700002642-11. 2. Label the vial as Gradient Performance Test. NOTE: There is no gradient performance test for the ACQUITY RI Detector.
Preparing for the system performance test 1. From Console, select ACQUITY UPLC System in the system tree, then Control > Start up system. a. For Prime Solvents (QSM tab), check all boxes and set “Duration of Prime” to 5 minutes. b. For Prime Solvents (SM tab), prime “Wash solvent” for 200 seconds and “Purge solvent” for 40 cycles. c. Click Start.
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2. Place a mobile phase blank (diluent) into the SM-FTN. 3. Place the appropriate test sample into the SM-FTN for the system precision test. 4. Place the component mix sample into the SM-FTN for the gradient performance test. 5. Flush the system to waste using a 25/25/25/25 mixture of mobile phases A, B, C, and D at 2 mL/min for 10 minutes.
Running maintenance tests NOTE:
Use the default parameter settings for tests in this section.
1. Run the Quaternary Solvent Manager Leak test using 100% acetonitrile (solvent B or D for TUV, PDA, ELS, FLR) or 100% RI Detector Qualification Mobile Phase (79.1%:20.9% water/methanol; RI only). When the test finishes, print the results. 2. Run the Sample Manager - FTN Calibrate needle z axis. Print the results. CAUTION:
TO PREVENT NEEDLE DAMAGE, DO NOT ALLOW THE TIP OF THE NEEDLE TO DIRECTLY
CONTACT THE BASE.
3. Run the Sample Manager - FTN Characterize needle seal test. Confirm that the test passes, then print the results. 4. Run the Sample Manager - FTN Leak test for the Sample syringe. Confirm that the test passes, then print the results.
Running a system leak test 1. Plug a V-detail pin plug into the V-detail union that is already attached to the APH assembly, then re-install the assembly into the column compartment. 2. Ensure that column temperature control is off. 3. From Console, select the Quaternary Solvent Manager from the system tree, click Maintain > Leak test using 100% acetonitrile (solvent B or D for TUV, PDA, ELS, FLR) or 100% RI Detector Qualification Mobile Phase (79.1%:20.9% water/methanol; RI only). 4. In the End Point section of the Parameters tab, click the Column option button, then select the Include Sample Manager Needle and Seal. 5. Click Start. Confirm that the test passes, then print the results.
Connecting and equilibrating the column 1. Remove the APH assembly from the column compartment. 2. Attach the outlet of the APH assembly to the inlet of the ACQUITY UPLC BEH C18 (2.1 x 50 mm, 1.7um, part number 186002350) column. To attach the APH to the column, refer to step 21 to step 24 on page 19 as appropriate for your system. CAUTION: THE DIRECTION OF THE FLOW IS CRITICAL. IF THE COLUMN IS INSTALLED IN THE WRONG DIRECTION, THE PACKING MATERIAL WILL BE DISTURBED, RESULTING IN LOSS OF EFFICIENCY AND POSSIBLE DAMAGE TO THE DETECTOR FLOW CELL.
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3. Plug the column with APH assembly into the column tray: a. Snap the column support clips, P/N 205000478, onto the body of the column (one between the reusable fitting and column). Rotate the clips so that the openings face forward.
Figure 25 - Column support clip installation b. Place the column in the column compartment. 4. Flush the column to waste with 100% acetonitrile (TUV, PDA, ELS, FLR) or 100% RI Detector Qualification Mobile Phase (79.1%:20.9% water/methanol; RI only) at 0.1 mL/ min for at least one hour before connecting the column outlet to the detector. 5. Attach the eCord. 6. Connect the column outlet to the detector. 7. Close the column heater door, and then inspect the seal around the front panel to ensure it aligns correctly. 8. For the TUV, PDA, ELS, FLR: a. Set the CH-A or CM-A temperature to 40° C and the Sample compartment to 10° C, then confirm that the temperatures are being regulated. b. From Console, set flow to 90%A:10%B at 0.6 mL/min and equilibrate until the baseline is stable (approximately 30 minutes). For the RI: a. Set the CH-A or CM-A temperature to 35° C and the Sample compartment temperature to OFF, then confirm that the temperatures are being regulated. b. From Console, set flow to 100% A at 0.4 mL/min and equilibrate until the baseline is stable (approximately 30 minutes) 9. While equilibrating, run the Sample Manager - FTN Needle Seal Readiness Test. Make sure that flow is set to 0.6 mL/min (TUV, PDA, ELS, FLR) or 0.4 mL/min (RI only). Confirm that the test passes, then print the results.
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Running the system precision test The instrument or inlet method you use to run the system precision test varies according to the type of detector (TUV, PDA, ELS, FLR, or RI) installed on the system. • • • • •
For TUV-based systems use: sys_prec_TUV For PDA-based systems use: sys_prec_PDA For ELS-based systems use: sys_prec_ELS For FLR-based systems use: sys_prec_FLR For RI-based systems use the instrument method parameters shown in Figure 34 through Figure 37.
NOTE:
The RI detector instrument method parameters (Figure 34 through Figure 37) detail the instrument method for an ACQUITY UPLC System. Use the same parameters for the ACQUITY H-Class System.
Checking the method parameters (TUV, PDA, ELS, FLR) 1. Open the instrument or inlet method appropriate for the system, then verify that the parameters settings are set up as shown in Figure 26 through Figure 33. Parameters for both Empower- and MassLynx-based systems must match. NOTE: The Quarternary Solvent Manager parameters are the same for the TUV, PDA, and ELSD detectors.
Figure 26 - System Precision Parameters for the Quarternary Solvent Manager for the TUV, PDA, or ELSD
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Figure 27 - System Precision Parameters for the Quarternary Solvent Manager for the TUV
Figure 28 - System Precision Parameters for the Quarternary Solvent Manager for the FLR
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NOTE: The Sample Manager - FTN parameters are the same for the TUV, PDA, and ELSD detectors (see Note about FLR detectors, below).
10/90 ACN/Water
Figure 29 - System Precision Parameters for the Sample Manager - FTN NOTE:
For systems with an FLR detector, Wash Solvent is 100% acetonitrile.
Figure 30 - System Precision Parameters for the PDA
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Figure 31 - System Precision Parameters for the TUV
Figure 32 - System Precision Parameters for the ELSD
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Figure 33 - System Precision Parameters for the FLR
Creating the method parameters (RI only)
Figure 34 - System Precision Parameters for the Quarternary Solvent Manager (RI only)
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)
Figure 35 - System Precision Parameters for the Sample Manager - FTN (RI only)
Figure 36 - System Precision Parameters for the (optional) Column Heater (RI only)
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)
Figure 37 - System Precision Parameters for the RI 2. For the TUV, PDA, ELS, or FLR detectors, open the appropriate sample set method for the system precision test and confirm the following: • • • •
A 30-minute equilibrate step that uses initial conditions One blank injection (TUV, PDA, ELS, FLR); (two blank injections for the RI) Six sample injections Injection volume = 5 µL (using 10-mm flow cells for PDA and TUV detectors); 1 µL (using 25-mm flow cells for PDA and TUV detectors); 5 µL for ELS, FLR, and RI detectors • Run time = 1.0 minute; 3.0 minutes (RI only) 3. Run the sample set. 4. When the run finishes, process the samples, then analyze the results and view the reports.
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System precision test results
273 nm
Figure 38 - System Precision Chromatogram for TUV and PDA (10-mm flow cell) NOTE:
If using the 25-mm flow cell, the peak area for the caffeine peak will be approximately 4x larger.
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Figure 39 - System Precision Chromatogram for ELSD
Figure 40 - System Precision Chromatogram for FLR
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Figure 41 - System Precision Chromatogram for RI 1. Verify that the peak is symmetrical, integrated, and identified correctly. 2. Standard deviation for peak retention over six replicate injections must be ≤ 1.0 second for caffeine and ≤ 2.8 seconds for anthracene. 3. If needed, follow these steps to use the Windows® scientific calculator: a. Click the Windows Start button and select Programs > Accessories > Calculator. The Windows calculator appears. b. Select Scientific from the View menu. The calculator switches to scientific view. c. Enter the first value by clicking the appropriate numbers on the Windows calculator (you can also use the numeric keypad to enter values if Num Lock is on). d. Click Sta in the left column of the calculator. The statistics box appears. e. Click Dat in the left column of the calculator, and then click Sta again to display the statistics box. Confirm that the statistics box displays the value entered in Step 3d. f. Enter each of the remaining values by clicking the appropriate numbers on the calculator, and then click Dat to enter each value in the statistics box. g. After you enter all values in the statistics box, click the s button in the left column of the calculator to calculate the standard deviation. 4. %RSD for peak areas over six replicate injections must be ≤ 0.5% (TUV and PDA), ≤ 5.0% (ELSD), ≤ 3.0% (FLR), and ≤ 1.5% (RI). 5. %RSD for peak heights over six replicate injections must be ≤ 0.9% (TUV and PDA), ≤ 5.0% (ELSD), and ≤ 4.5% (FLR). NOTE: %RSD for peak heights is not calculated for the ACQUITY RI Detector.
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6. Record the results in the appropriate tables in the Waters ACQUITY UPLC H-Class System with TUV, PDA, ELS, FLR, or RI Detectors Installation Checklist.
Running the gradient performance test The instrument or inlet methods you use to run the gradient tests vary according to the type of detector (TUV or PDA) installed on the system. • For TUV-based systems use: gradient_AB_TUV and gradient_CD_TUV • For PDA-based systems use: gradient_AB_PDA and gradient_CD_PDA NOTE:
The Gradient Performance Test is not performed for the ACQUITY ELS, FLR, or RI detectors.
1. Open the instrument or inlet method appropriate for the system (start with the method for solvents A and B), then verify that the parameters settings are set up as shown in Figure 42 through Figure 47. Parameters for both Empower- and MassLynx-based systems must match. NOTE: Use the same Quarternary Solvent Manager parameters for the TUV or the PDA.
Figure 42 - Gradient Performance Parameters for the Quarternary Solvent Manager (solvents A and B; rows 1 to 4)
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Figure 43 - Gradient Performance Parameters for the Quarternary Solvent Manager (solvents A and B; rows 3 to 6)
Figure 44 - Misc Tab – Gradient Performance Parameters for the Quarternary Solvent Manager
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NOTE: Use the same Sample Manager parameters for the TUV or the PDA.
10/90 ACN/water
Water
Figure 45 - Gradient Performance Parameters for the Sample Manager
20
Figure 46 - Gradient Performance Parameters for the PDA
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Figure 47 - Gradient Performance Parameters for the TUV 2. Open the appropriate sample set method for the gradient test and confirm the following settings are in place: • • • •
A 6-minute condition column step that uses initial conditions Three sample injections Injection volume = 5 µL (using 10-mm or 25-mm flow cells) Run time = 4.0 minutes
3. Run the sample set. 4. When the run finishes, process the samples, then analyze the results and view the reports.
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Gradient performance test results
Solvents A and B
Solvents C and D
Figure 48 - Gradient Performance Chromatograms for AB and CD NOTE:
If using the 25-mm flow cell, the peak areas will be approximately 4x larger.
1. Verify that the peaks are symmetrical, integrated, and identified correctly. 2. The standard deviation for the peak retention over three replicate injections must be ≤ 1.5 seconds. 3. Repeat the gradient performance tests that start on page 39 for mobile phases C and D. 4. Record the results in the appropriate table in the Waters ACQUITY UPLC H-Class System with TUV, PDA, ELS, FLR, or RI Detectors Installation Checklist.
